Dead Cell Apoptosis Kits with Annexin V for Flow Cytometry
Dead Cell Apoptosis Kits with Annexin V for Flow Cytometry
Invitrogen™

Dead Cell Apoptosis Kits with Annexin V for Flow Cytometry

Assess cell viability with Dead Cell Apoptosis kits with Annexin V and conjugated fluorescent dyes such as Alexa Fluor 488, FITC, PI, PE, APC, and SYTOX Green.
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Catalog NumberQuantityLabel or Dye
V132421 KitFITC, Propidium Iodide
V1324150 AssaysAlexa Fluor 488, Propidium Iodide
V351121 KitSYTOX Green, R-PE
V351131 KitSYTOX Green, APC
V13245250 AssaysAlexa Fluor 488, Propidium Iodide
Catalog number V13242
Price (USD)
367.00
1 kit
Add to cart
Quantity:
1 Kit
Label or Dye:
FITC, Propidium Iodide
Price (USD)
367.00
1 kit
Add to cart
Easily differentiate live, dead, and apoptotic cells during flow cytometry with our Dead Cell Apoptosis kits with Annexin V conjugates including Alexa Fluor 488, FITC, propidium iodide, PE, APC, and SYTOX Green. These conjugates distinguish live, dead, or apoptotic cells via different dyes, which is vital for confirming cell viability and apoptosis using multi-parametric studies.
There are several advantages to using Dead Cell Apoptosis kits with Annexin V for flow cytometry in order to assay cell viability:

Superior brightness
Unlike other Annexin V kits that have lower protein concentrations or purification levels, the Annexin V Alexa Fluor 488 conjugate is optimized for flow cytometry to provide the largest separation between apoptotic and live cells. The Alexa Fluor 488 dye is a superior green-fluorescent dye with a spectrum similar to fluorescein (FITC).

High binding efficiency
Annexin V conjugates are made from a highly purified cys-annexin, resulting in higher binding efficiency and extremely accurate characterization of the apoptotic process.

Multi-parametric
Many publications require a minimum of two different ways to identify apoptotic cells. This multi-parametric kit detects phosphatidyl serine (PS) on the cytoplasmic surface of the cell membrane, while membrane integrity is determined via propidium iodide.

Dead Cell Apoptosis Kit with Annexin V Alexa Fluor 488 & PI
The Dead Cell Apoptosis Kit with Annexin V Alexa Fluor 488 & Propidium Iodide (PI) (V13241, V13245) is used in flow cytometry to measure early apoptosis by detecting phosphatidyl serine (PS) expression and membrane permeability. When cells are stained with Annexin V and propidium iodide, apoptotic cells expressing PS show green fluorescence, which can be detected in the FITC channel, and low red fluorescence. Dead or necrotic cells show bright red fluorescence and no green fluorescence, while live cells show no green or red fluorescence.

Dead Cell Apoptosis Kit with Annexin V PE and SYTOX Green
The Dead Cell Apoptosis Kit with Annexin V PE and SYTOX Green detects PS externalization in apoptotic cells during flow cytometry using recombinant annexin V conjugated to the orange fluorescent phycobiliprotein R-PE, while dead cells are detected with SYTOX Green nucleic acid stain. After treatment with both probes, apoptotic cells show orange fluorescence, dead cells show green fluorescence, and live cells show little or no fluorescence. These populations can easily be distinguished in the 530/30 nm and 585/42 nm bandpass filters with a 488 nm laser flow cytometer.

Dead Cell Apoptosis Kit with Annexin V APC and SYTOX Green
The Dead Cell Apoptosis Kit with Annexin V APC and SYTOX Green detects PS externalization in apoptotic cells during flow cytometry using recombinant annexin V conjugated to red laser-excited allophycocyanin, and dead cells using SYTOX Green nucleic acid stain. Apoptotic cells are detected by annexin V binding to externalized PS. Live cells show little or no fluorescence, apoptotic cells show red fluorescence and very little green fluorescence, and late apoptotic cells show a higher level of red and orange fluorescence. These populations can easily be distinguished using a flow cytometer that has both 488 nm and 633 nm excitation sources (an argon-ion laser and a HeNe laser).

The Dead Cell Apoptosis Kit with Annexin V FITC and PI
The Dead Cell Apoptosis Kit with Annexin V FITC and PI detects PS externalization in apoptotic cells using recombinant annexin V conjugated to green-fluorescent FITC dye and dead cells using PI. Necrotic cells that stain with PI show red fluorescence. After treatment with both probes, apoptotic cells show green fluorescence, dead cells show red and green fluorescence, and live cells show little or no fluorescence.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
DescriptionDead Cell Apoptosis Kit with Annexin V FITC and PI, for flow cytometry
Excitation/Emission494, 535/518, 617
Flow Cytometer Laser Lines488
For Use With (Equipment)Flow Cytometer
Kit ContentsContains 1 vial of annexin V, FITC conjugate (250 μL ), 1 vial of propidium iodide (PI, 100 μL), and 1 bottle of annexin binding buffer (5X solution, 15 mL).
Label or DyeFITC, Propidium Iodide
No. of Reactions50
Product TypeApoptosis Detection Kit
Quantity1 Kit
Shipping ConditionWet Ice
Unit Size1 kit
Contents & Storage
Store in refrigerator (2°C to 8°C) and protect from light.
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Frequently asked questions (FAQs)

I want to study apoptosis using an Annexin V conjugate, but with adherent cells via microscopy instead of flow cytometry. Can this be done?

It has been done, but we don‘t recommend it. Both healthy cells and apoptotic cells possess phosphatidylserine on the cell surface, which can be detected with Annexin V, but apoptotic cells have significantly more of it. You can easily tell the difference between these two populations with flow cytometry, because flow cytometers are more sensitive and have a higher throughput. But with a microscope, you cannot always tell the difference, especially for adherent cells. Instead, for microscopy, we recommend a different technique, such as detecting caspases with CellEvent Caspase Detection Reagents.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

When using Dead Cell Apoptosis Kits with Annexin V for Flow Cytometry, what does a PI+ only population (hence Annexin V negative) correspond to?

These products are cells that died without undergoing apoptosis. A PI+ only population could be either a result of experimental condition or sample preparation.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I trypsinized my adherent cells and labeled with annexin V, and now my flow data is showing a high percentage of apoptotic cells even for control, untreated cells. What is the problem?

Trypsinization or mechanical scraping of cells temporarily disrupts the plasma membrane, allowing annexin V to bind phosphatidylserine on the cytoplasmic surface of the cell membrane and thus leading to false positive staining. Allow the cells to recover for about 30 minutes in optimal cell culture conditions and medium after trypsinizing/scraping so that they can recover their membrane integrity before staining. For lightly adherent cell lines, such as HeLa and NIH 3T3, another option is to use non-enzyme treatments like Gibco Cell Dissociation Buffer (Cat. No. 13151014).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Can I detect annexin V staining in an imaging assay?

Annexin V staining is not typically used in imaging experiments; it is a better reagent for flow cytometry analysis. All cells will stain to some extent, so it can be difficult to distinguish a relatively bright annexin V-stained cell from a dimmer non-apoptotic cell. Caspase activation, detected using our CellEvent Caspase 3/7 or Image-iT LIVE Caspase detection kits, is a better method for detecting apoptosis in an imaging assay.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

When should I stain adherent cells with annexin V for flow cytometric analysis? Before or after I trypsinize them?

Trypsinize first and then allow the cells to recover about 30 minutes in optimal cell culture conditions and medium before staining with annexin V conjugates. Trypsinization or mechanical scraping of cells temporarily disrupts the plasma membrane, allowing for annexin V to bind phosphatidylserine on the cytoplasmic surface of the cell membrane and thus leading to false positive staining. For lightly adherent cell lines such as HeLa and NIH 3T3, you could use a less harsh (non-enzymatic) dissociation product like Gibco Cell Dissociation Buffer (Cat. No. 13151014).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

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